Genome editing in plants by engineered CRISPR-Cas9 recognizing NG PAM

Nat Plants. 2019 Jan;5(1):14-17. doi: 10.1038/s41477-018-0321-8. Epub 2018 Dec 10.

Abstract

Streptococcus pyogenes Cas9 (SpCas9) is widely used for genome editing and requires NGG as a protospacer adjacent motif (PAM). Here, we show that the engineered SpCas9 (SpCas9-NGv1) can efficiently mutagenize endogenous target sites with NG PAMs in the rice and Arabidopsis genomes. Furthermore, we demonstrate that the SpCas9-NGv1 nickase fused to cytidine deaminase mediates C-to-T substitutions near the 5' end of the target sequence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics*
  • CRISPR-Associated Protein 9 / genetics*
  • CRISPR-Cas Systems*
  • Gene Editing / methods*
  • Genome, Plant
  • Nucleotide Motifs
  • Oryza / genetics*
  • Plants, Genetically Modified / genetics
  • RNA, Guide, CRISPR-Cas Systems
  • Transformation, Genetic

Substances

  • RNA, Guide, CRISPR-Cas Systems
  • CRISPR-Associated Protein 9
  • Cas9 endonuclease Streptococcus pyogenes